Human Phospho-GSK-3 beta (S9) Antibody Summary
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human Phospho-GSK‑3 beta (S9) by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nM PMA for 20 minutes. PVDF Membrane was probed with 1 µg/mL of Human Phospho-GSK-3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-GSK-3 beta (S9) at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-GSK‑3 beta (S9) in HeLa Human Cell Line. GSK-3 beta phosphorylated at S9 was detected in immersion fixed HeLa human cervical epithelial carcinoma cells, unstimulated (lower panel) or stimulated (upper panel) with PMA, using Mouse Anti-Human Phospho-GSK-3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Phospho-GSK-3 beta (S9) in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line untreated (open histogram) or treated with 120 ng/mL PMA for 15 minutes (filled histogram) was stained with Human Phospho-GSK-3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) or isotype control antibody (Catalog # MAB002, not shown), followed by Fluorescein-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0103B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: GSK-3 beta
Glycogen Synthase Kinase-3 (GSK-3) is a serine/threonine kinase initially identified asan inhibitor of glycogen synthase. Two isoforms (GSK-3 alpha and GSK-3 beta ) share 85% amino acid identity. GSK-3 beta, inhibited by phosphorylation at S9 by Akt, is involved in energy metabolism, body pattern formation, and neuronal cell development.
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